Department of Biosciences, School of Natural Sciences, Technical University of Munich, Am Coulombwall 4a, 85748 Garching, Germany.
Munich Institute of Biomedical Engineering, Technical University of Munich, Boltzmannstraße 11, 85748 Garching, Germany.
J Am Chem Soc. 2023 Mar 8;145(9):4946-4950. doi: 10.1021/jacs.2c12733. Epub 2023 Feb 24.
DNA origami enables the creation of complex 3D shapes from genetic material. Future uses could include the delivery of genetic instructions to cells, but nuclear import remains a major barrier to gene delivery due to the impermeability of the nuclear membrane. Here we realize active nuclear import of DNA origami objects in dividing and chemically arrested mammalian cells. We developed a custom DNA origami single-strand scaffold featuring a mammalian-cell expressible reporter gene (mCherry) and multiple Simian virus 40 (SV40) derived DNA nuclear targeting sequences (DTS). Inclusion of the DTS within DNA origami rescued gene expression in arrested cells, indicating that active transport into the nucleus occurs. Our work successfully adapts mechanisms known from viruses to promote the cellular expression of genetic instructions encoded within DNA origami objects.
DNA 折纸术可利用遗传物质创建复杂的 3D 形状。未来的用途可能包括向细胞输送遗传指令，但由于核膜的不渗透性，核内输入仍然是基因输送的主要障碍。在这里，我们在分裂和化学固定的哺乳动物细胞中实现了 DNA 折纸物体的主动核内输入。我们开发了一种定制的 DNA 折纸单链支架，具有哺乳动物细胞可表达的报告基因（mCherry）和多个猴病毒 40（SV40）衍生的 DNA 核靶向序列（DTS）。在 DNA 折纸术内包含 DTS 可挽救被阻止的细胞中的基因表达，表明发生了主动运输到细胞核内。我们的工作成功地利用了已知来自病毒的机制来促进 DNA 折纸物体中编码的遗传指令的细胞表达。
